Volume 270,
Number 2,
Issue of January 13, 1995 pp. 849-853
©1995 by The American Society for Biochemistry and Molecular Biology, Inc.
Inhibition
of Zona Pellucida Gene Expression by Antisense Oligonucleotides
Injected Into Mouse Oocytes
(Received for publication, September 15, 1994; and in revised form, November 1, 1994)
Zhi-Bin
Tong
,
Lawrence M.
Nelson
,
Jurrien
Dean
During murine oogenesis, the zona pellucida proteins (ZP1, ZP2,
and ZP3) are synthesized and secreted to form an extracellular matrix
that surrounds the oocyte and mediates specific biological functions
essential to mammalian fertilization and early development. To
investigate the relationship among the zona proteins during zona matrix
assembly, we have undertaken to inhibit de novo biosynthesis
of specific zona proteins with antisense oligonucleotides complementary
to the 5`-ends of ZP2 (nucleotide position 19-42) and ZP3
(nucleotide 21-44) mRNAs. When injected into the cytoplasm of
growing mouse oocytes, the antisense oligonucleotides targeted specific
zona mRNAs for degradation, as confirmed by a RNase protection assay.
Individual zona pellucida protein synthesis was followed by
immunoprecipitation with ZP2- and ZP3-specific monoclonal antibodies.
New zona protein synthesis from the targeted mRNA was abolished, but
nontargeted zona protein continued to be synthesized. Interestingly,
abolishment of either ZP2 or ZP3 protein synthesis prevented the
incorporation of the other protein into the extracellular zona matrix.
These results suggest that ZP2 and ZP3 proteins are independent of each
other in their biosynthesis but are dependent upon each other for their
incorporation into the zona pellucida matrix. This study provides an
experimental system in which destruction of a targeted mRNA generates a
transient loss-of-expression phenotype during mouse oocyte growth.
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